Impact of basic soil parameters on pesticide disappearance investigated by multivariate partial least square regression and statistics.

Dissipation time is a key parameter when studying and modeling the environmental fate of pesticides. This study was conducted to characterize the variability of pesticide disappearance in soil and to identify possible controlling parameters related to intrinsic soil properties and microbiology. Multivariate data analysis was used to study spatial variability in three horizons from 24 sandy soil profiles. The time for 50% disappearance (DT(50)) was characterized for two herbicides, metribuzin (MBZ) and MCPA, and methyltriazine amine (MTA; transformation product of metsulfuron-methyl, tribenuron-methyl, thifensulfuron-methyl, and chlorsulfuron). Normal and log-normal distributions were compared for DT(50) and soil properties and descriptive statistics were calculated. Conformity with log-transformed distributions was observed and assuming normality of the DT(50) data would cause 5 to 35% overestimation. Mean DT(50) were: MCPA 9.5, MBZ 168, and MTA 127. Significant effect of soil depth on DT(50) was shown for MCPA and MBZ, with low values in deeper horizons. Simple linear correlation for combinations of MCPA, MTA, pH, and total organic carbon (TOC) was observed. Using partial least squares regression (PLS) 71 to 85% of the total DT(50) variance was explained. A specific predictor variable could not be identified as the controlling components differed within horizons and compounds. For MCPA the overall important predictor variables were microbiology and TOC, whereas for MTA and MBZ it was inorganic variables (Al, Fe, cation exchange capacity, base saturation percent, and pH) and microbiology. The study indicates that PLS generated input data can improve pesticide fate modeling and reduce the uncertainty in dissipation estimation.

Analysis of acidic pesticides using in situ derivatization with alkylchloroformate and solid-phase microextraction (SPME) for GC-MS.

A solid-phase microextraction (SPME) method was developed for the analysis of acidic pesticide residues in water. The method utilizes in situ derivatization with butylchloroformate (BuCF), followed by on-line SPME extraction using a PDMS fibre, and analysis by GC-MS. Derivatives of the phenoxy acids mechlorprop (MCPP), dichlorprop (DCPP), MCPA and 2,4-D and their phenol degradation products 4-chloro-2-methylphenol and 2,4-dichlorophenol (DCP) were identified. Detection limits at 0.16-2.3 microg/l were achieved. Optimization of derivatization, ion strength, extraction time, SPME-fibre, desorption time and temperature are described. Standard curves in the range 0.5-10.0 microg/l were fitted to a second-degree polynomial. Standard deviation (n = 5) was below 10% for the phenol derivatives, but 20-50% for the phenoxy acids. For method verification groundwater samples from a field experiment were screened for content of MCPP and compared to the results from the HPLC analysis. A good agreement was obtained with respect to identification of positive samples, even though concentrations measured by the SPME were lower than with HPLC. Even if the precision and accuracy do not meet the demands for a strictly quantitative analysis, the SPME method is suitable for screening, because it is cheap, it can be automated, and uses smaller amounts of potential harmful solvents. Also, the method is less labour-intensive, as it requires a minimum of sample preparation when compared to traditional analyses. The acidic pesticides bentazon, dicamba, bromoxynil, ioxynil, dinoseb and DNOC were included in the study but could not be analysed by the current method.

Analysing transformation products of herbicide residues in environmental samples.

High-performance liquid chromatography (HPLC) methods were developed for the optimised determination of five herbicide residues (dichlorprop, isoproturon, mecoprop, metsulfuron-methyl and 2,4,5-T) and major metabolites. These compounds represent important groups of herbicides and several residues have been found as contaminants in groundwater. The methods make it possible to study these herbicides and several transformation products through simultaneous detection and quantification. Culture media as well as cleaned up extracts from sediment and groundwater can be analysed. Using HPLC with UV detection the general limit of quantification was 1.8 ng injected corresponding to a detection limit of 1-2 micrograms/l when analysing a cleaned up extract from a 20 ml water sample. The method was verified by analysing herbicide residues in groundwater collected from a wetland area. Cleaning up 20 ml groundwater with a residue level of 25 micrograms/l the general recovery was within 58-82%.

Human semen quality in relation to dietary pesticide exposure and organic diet.

The objective of the study was to corroborate or refute the hypothesis that farmers having a high intake of organic grown commodities have a high semen quality due to their expected lower level of dietary pesticides intake. Food frequency data and semen were collected from 256 farmers (171 traditional farmers and 85 organic farmers, overall participation rate: 32%) who were selected from central registers. Each farmer delivered one semen sample before the spraying season started. The farmers were divided into three groups where the commodities from organic production contributed no (N, 0%), medium (M, 1-49%), or a high (H, 50-100%) proportion of the fruit and vegetables consumed. Farmers having a high relative intake of organically grown fruit and vegetables also had a high relative consumption of organically produced meat, milk, and bread, and differences were observed comparing the actual mean intake of single commodities, such as rice, potato, and pork meat. The current individual dietary intake of 40 pesticides was estimated using food frequencies and generalized serving size data in combination with data on pesticide concentrations in food commodities as obtained from the National Danish Food Monitoring Program. The estimated pesticide intake was significantly lower among farmers of group H, but for all three groups of farmers the average dietary intake of 40 pesticides was at or below 1% of the acceptable daily intake (ADI) except for the dithiocarbamates (max = 0.21 microg/kg day = 2.2% ADI), methidathion, (max = 0.01 microg/kg day = 1.4% ADI), and 2-phenylphenol (max = 0.21 microg/kg day = 1.1% ADI). The median sperm concentration for the three groups of farmers was not significantly different (p = 0.40, median sperm concentration was N = 62, M = 44, and H = 75 million/ml). The group of men without organic food intake had a significant lower proportion of morphologically normal spermatozoa, but in relation to 14 other semen parameters no significant differences were found between the groups. Intake of 40 individual pesticides was correlated with four semen parameters (concentration, percentage dead spermatozoa, percentage normal sperm heads, and motility [VCL]). Five significant correlations (p value 0.01) were found among the 160 comparisons in relation to percentage dead spermatozoa: azinphos-methyl, carbaryl, chlorfenson, fenitrothion, and tetradifon. For all of them a lower percentage of dead spermatozoa were found in the groups with a high dietary intake of the specific pesticide. In contrast, for all pesticides evaluated only minor differences were found between the groups when considering spermatozoa concentration, morphology, and motility. In conclusion, the estimated dietary intake of 40 pesticides did not entail a risk of impaired semen quality, but precautions should be taken when generalizing this negative result to populations with a higher dietary exposure level or an intake of other groups of pesticides.

Residues of chlormequat and mepiquat in grain--results from the Danish National Pesticide Survey.

The objective of the present work was to establish information on chlormequat and mepiquat residues in grain for human consumption. Chlormequat (2-chloro-N,N,N-trimethylethylammonium, CAS RN 7003-89-6) and mepiquat (1,1-dimethylpiperidinium, CAS RN 15302-91-7) are plant growth regulators used to stabilize stalks in cereals. The study was part of the Danish National Pesticide Survey, managed by the Danish Veterinary and Food Administration. Samples were collected in autumn 1997. Residue contents were determined with a newly developed liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for chlormequat analysis. The method was extended to include mepiquat in the present study. Quantitation was done by the internal standards method, using mass chromatograms of the most intense daughter ions of mepiquat (m/z 98), chlormequat (m/z 58), and [13C]-chlormequat (m/z 61, internal standard). For chlormequat, the overall limit of detection (LD) was 6 micrograms/kg and the limit of determination (LOD) was 10 micrograms/kg. For mepiquat, LD was 2 micrograms/kg and LOD was 3 micrograms/kg. Of 77 samples analyzed, 51 contained chlormequat and 11 contained mepiquat. The highest levels of chlormequat were found in samples of oatmeal (3.76 mg/kg) and rye (1.08 mg/kg). In 9 rye grain samples containing chlormequat, 5 also contained mepiquat. However, in all samples analyzed, the residues of chlormequat and mepiquat were below maximum residue limits.

Pesticide residues in selected food commodities: results from the Danish National Pesticide Monitoring Program 1995-1996.

The paper presents results of analyses of 4182 samples collected from January 1995 to December 1996 for the Danish National Pesticide Monitoring Program. The program basis is a random sample control supplemented with a target control. The objectives of the control are to monitor consumer exposure to health hazards and to enforce national and European Community regulations. The Danish Veterinary and Food Administration is responsible for establishing methods, data manipulation, and evaluation, but actual analyses are performed at 4 regional laboratories. In addition to a description of the Danish National Pesticide Monitoring Program on food, the effects of lowering the reporting limits are discussed. Pesticides included are those in current use as well as chlorinated pesticides like lindane, DDT, and HCB, occurring in food now primarily as a result of environmental contamination. Commodities analyzed are fruits, vegetables, cereals, bran, fish, and animal products such as meat, butter, cheese, fat, and eggs. In fruits and vegetables, residues were detected in 10% of 2515 samples, with higher incidences of detection for foreign commodities. Violation rate was 0.6%. In food of animal origin, low levels of organochlorine pesticides were detected in most fish samples and in more than half of the animal product samples. However, no results exceeding maximum residue limits were found. No residues of organophosphorus pesticides were detected in the 231 meat samples analyzed.

Supercritical fluid extraction of pesticides from meat: a systematic approach for optimisation.

A method for quantification of pesticide residues in meat and fatty matrices was developed using supercritical fluid extraction (SFE). The SFE method allows selective extraction of residues and subsequent gas chromatography analysis without further clean-up. Quantification was done by GC using nitrogen-phosphorus detection and electron capture detection. Initial method development was made using organophosphorus pesticides (OPPs). The dependence of fat and OPP residue recovery on supercritical fluid density, temperature, flow rate and extraction time was investigated through a reduced factorial design. Since temperature and density were found to have pronounced effect on the recovery of OPPs these extraction parameters were studied using a new arbitrary measure for co-extractivity. An optimisation score was established as relative pesticide recovery subtracted by relative fat recovery. Using this algorithm a response plane was modelled varying the primary factors temperature and density. The applicability of this approach and the algorithm was verified. The polarity range covered by the SFE method was demonstrated using OPPs: chlorpyrifos, chlorpyrifos-methyl, malathion, pirimifos-methyl and prothiofos. Additionally the final method was evaluated using four pesticides that are not OPPs: carbofuran, phorate, procymidone and vinclozolin. All pesticides showed good recovery (78-95%), and limits of detection (0.01-0.03 mg kg-1) and limits of determination (0.01-0.05 mg kg-1) meet the requirements set by the European Council (Directive 96/33/EEC). Compared to traditional methods based on organic solvent extraction, the SFE method is fast, less labour intensive, uses smaller amounts of potentially harmful solvents and has the potential to be fully automated.

Optimized method for the determination of organophosphorus pesticides in meat and fatty matrices.

A method was developed and optimized for determination of residues of organophosphorous pesticides (OPs) in meat and fatty matrices. The method was developed for the national Danish monitoring programme, whereby priority was given to simplify the clean-up, avoid use of toxic and harmful organic solvents and allow quantification at ppb level using GC with nitrogen-phosphorus detection (NPD). Homogenized meat was extracted using ethyl acetate, the co-extracted water being removed using anhydrous Na2SO4. The clean-up was done through fat precipitation by cooling the extract flowed by solid-phase extraction on C18 mini-columns. Pesticide residues were determined by GC-NPD using a DB1701 capillary column. The limit of detection was 1 to 20 ppb and limit of determination was 2 to 33 ppb and the method is feasible for control of fat-soluble OPs according to the maximum residue limits set by the European Communities. The method developed covers a broad polarity range from polar OPs, such as acephate and methamidophos, through medium polar OPs to non-polar pesticides, such as prothiofos.

Composition of photosynthetic pigments in thylakoid membrane vesicles from spinach.

Thylakoid membranes from spinach were fragmented mechanically and separated into vesicles originating from grana and stroma-exposed lamellae (Andreasson et al. (1988) Biochim Biophys Acta 936: 339-350). The grana vesicles were further fragmented and separated into smaller vesicles originating from different parts of the grana (Svensson and Albertsson (1989) Photosynth Res 20: 249-259). All vesicles so obtained were analyzed with respect to chlorophyll and carotenoid composition by reverse phase HPLC. For all fractions the following relations (mole/mole) were found: 1 carotenoid per 4 chlorophyll (a+b), 2 lutein per 5 chlorophyll b and 5 violaxanthin per 100 chlorophyll (a + b). The contents of lutein and neoxanthin were each linearly related to chlorophyll b and ß-carotene was linearly related to chlorophyll a.

Chlorophyll fluorescence transients in a barley mutant lacking Photosystem I.

We have compared the properties of a mutant of barley lacking Photosystem I (viridis-zb (63) ) with the corresponding wild type using modulated fluorescence measurements. The mutant showed two unexpected characteristics. Firstly, there was a slow decline in the fluorescence signal in the light which was dependent on the presence of O2 at concentrations similar to that in air; 2% O2 in N2 had no effect. The observed decline was mainly due to an increase in the non-photochemical quenching. Secondly, in the absence of O2, saturating light pulses caused a pronounced transient decrease in the fluorescence signal; a similar effect could also be observed in wild type plants when neither CO2 nor O2 was present.